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. 2021 Apr;191(4):590–601. doi: 10.1016/j.ajpath.2021.01.001

Figure 4.

Figure 4

Intestinal cells express the tissue-type plasminogen activator (tPA) cell-signaling response system. A: Flow cytometry was performed to detect the glutamate ionotropic receptor NMDA type subunit 1 (GluN1) N-methyl-d-aspartate receptor (NMDA-R) subunit in CD45-positive cells isolated from mice exposed to 4% dextran sulfate sodium (DSS) for 5 days and then allowed to recover for 7 days. The mice were treated on day 5 with enzymatically inactive tPA (EI-tPA; DSS + EI-tPA; blue tracing) or with vehicle (DSS; orange tracing). Cells isolated from control (Con) mice that were not DSS-treated are labeled Control (black tracing). GluN1 immunofluorescence is compared with cells stained with secondary antibody only (II Ab only). B: The results of six separate flow cytometry studies are summarized. C: Flow cytometry was performed to detect the GluN1 NMDA-R subunit in epithelial cell adhesion molecule (EpCAM)–positive cells isolated from mice exposed to 4% DSS for 5 days and then allowed to recover for 7 days. The mice were treated with EI-tPA (DSS + EI-tPA; blue tracing) or with vehicle (DSS; orange tracing). Cells isolated from control mice that were not DSS-treated are labeled Control (black tracing). GluN1 immunofluorescence is compared with cells that were treated with II Ab only. D: The results of six separate flow cytometry studies are summarized. E: Flow cytometry was performed to detect the GluN1 NMDA-R subunit in CD45-positive cells expressing CD3, CD103, or Ly6G isolated from mice exposed to 4% DSS for 5 days (green tracing; representative of six separate studies). The secondary antibody only control is shown with shaded curves. F: Colons were harvested from control mice that were not DSS-exposed and from mice treated with 4% DSS for 5 days, treated with EI-tPA or vehicle, and allowed to recover for 7 days. Immunohistochemistry for low-density lipoprotein receptor–related protein-1 was performed. Data are presented as means ± SEM (B and D). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗∗P < 0.0001. Scale bars = 50 μm (F).